Treatment of a sample with focused acoustic energy

ABSTRACT

A device for treating a sample with focused acoustic energy is provided. The device transmits the generated acoustic energy from the source to the sample via a complete dry propagation path. A cartridge containing the sample is inserted into an instrument, wherein the insertion forms a dry propagation path.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International Application No.PCT/CH2010/000093 filed Apr. 9, 2010, now pending, which claims thebenefit under 35 U.S.C. §119(a) of European Patent Application No.09157850.0, filed Apr. 14, 2009, the entire contents of both of whichare incorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to the treatment of samples with focusedacoustic energy. In particular the invention relates to a device forirradiating a sample with focused acoustic energy to treat the sampleand a method for irradiating a sample with focused acoustic energy totreat the sample.

2. Description of Related Art

In recent years progress in many aspects of sample-in result-outdevices, also known as micro total analysis systems (microTAS) orlab-on-a-chip, has generated, for a variety of reasons, an increasinginterest in in-vitro-diagnostic (IVD) applications. For example theintegration and miniaturization results in systems requiring a relativesmall, acceptable contamination risk of the sample, high sensitivity andshort turn-around time of the test and lower costs per test. Furthermorebetween sample-input and result generation minimal operator interventionshall be required. Operator interventions can be done by relativelyunskilled operators and moderate demands on operating environment.

Known technologies of treating samples with acoustic energy may not beappropriate for certain applications like molecular device applicationsbecause after completion of the sonic treatment no distinction may bemade between a leaking cartridge having a liquid sample inside and theliquid being used by the device itself. That may be a non-acceptabletreatment result within these devices using such a liquid-coupling.

Furthermore the pretreatment function including complex operations likee.g. mixing, is processed separately and independently from otherprocessing functions. This is opposite to the general trend in thisdomain of further miniaturization and integration. Even more seriously,it contradicts with for example hospital or lab requirements to havereal small size systems, because of the very limited space available inthese settings.

In addition to that molecular diagnostic tests often includetechnologies with complicated piezo arrays, complicated control systemsand complicated electric drivers. These technologies are expensive,require a lot of technical support and also need much space.

SUMMARY OF THE INVENTION

It may be an object of the invention to provide for an improvedtreatment of samples.

Definitions and Abbreviations

It shall be noted that in the context of this invention the followingdefinitions and abbreviations will be used:

Dry coupling: The term “dry coupling” will be used in the context of theinvention as a complete transmission of the acoustic energy through onlynon-liquid matter from the source to the sample.

Acoustic energy: The term “acoustic energy” is in the context of theinvention used as comprising such terms as sonic energy, acoustic waves,acoustic pulses, ultrasonic energy, ultrasonic waves, ultrasound,Shockwaves, sound energy, sound waves, sonic pulses, pulses, waves orany other grammatical form of these terms.

Focal region and focal point: “Focal region” or “focal point” as used inthe context of the present invention means that a region where theacoustic energy converges and/or hits a target or sample, although thatregion has not necessarily to be a single focused point.

Device: The expression “device” in the context of the invention includesmolecular diagnostic devices as well as other devices. Applications ofthe device may e.g. be in healthcare/life science, food industry,veterinary practice and forensic applications.

Sample: It shall explicitly be noted that the term “sample” may containsamples for molecular analysis being treated with the device accordingto the present invention. For example blood, cultured blood, urine,aspirate, samples with water like viscosity, heterogeneous samples orsamples on a carrier like BAL, sputum, tracheal aspirate, CSF, swaband/or brush with pathogen. Nevertheless this does not mean that anyother kind of matter, solid, liquid, gaseous or any combination thereofis excluded from being a sample and being irradiated with focusedacoustic energy by the invention.

NA: “NA” will be used for any nucleic acid.

Source: In the context of the invention the term “source” will be usedsynonymously to the term transducer. Additionally any other apparatusthat is able to emit acoustic energy as defined within the context ofthe invention is comprised in the source.

Propagation path: The expression “propagation path” describes in thecontext of the invention the way of the acoustic energy from the sourcethrough any combination of at least the coupler and the cartridge to thesample. Other elements like lenses, additional couplers may be in thepropagation path. Thus in the propagation path also the intermediatecontact layers of these different elements are passed by the acousticenergy. Additionally other layers like e.g. the acoustic window or theinterface medium may be comprised.

Attenuation: The term “attenuation” in the context of the inventionrelates to a decrease of the intensity of the generated acoustic energy.This may be e.g. due to reflection, absorption, diffraction, or anycombination thereof.

Treatment of the sample: The term “treatment” or “treating” is used inthe context of the invention to describe the interaction of the focusedacoustic energy with the sample. By means of focusing the acousticenergy onto the sample in various specific ways sonochemical and/orsonophysical reactions are caused in the sample to generatefunctionalities like e.g. mixing, dispersing, stirring, elution fromswabs or brushes, liquefaction, lysing or cell release. Thereby thisdefinition of “treatment” also describes the sonophysical and/or thesonochemical interactions during the process called “pretreatment.” Inother words “treatment” comprises amongst other functionalities the“pretreatment” of a sample.

Process chamber: The expression “process chamber” will be similarly usedto “chamber” and “chamber of the cartridge.”

Ultrasound: The terms “ultrasound” and “ultrasonic” will be used forcyclic sound pressure with a frequency between 20 kHz and 100 kHz.

High Intensity Focused Ultrasound (HiFu): The term “HiFu” will be usedin the context of the invention as focused acoustic field with sourcefrequencies in the range of 0.2 MHz to 10 Mhz, with amplitudes chosen tobe sufficient efficient to create high pressure shock-waves and/orcavitation in the focal zone. Focal zone dimensions (length anddiameter) are dependent on the source transducer type (e.g. naturalfocusing by flat or enforced focusing by conical/spherical sourcetransducers). Exemplary length-scales for the indicated frequency rangeare (sub) millimeters.

Sample-in result out-system: A system which accepts a (e.g. biological)sample, does all the required preparation steps to prepare for detectingany kind of facts, runs the detection and delivers the detectionresults. For example a device for molecular analysis of samples likee.g. blood or other cells can be provided, that provides for allnecessary analysis steps from the supply of the natural, untreatedsample to the result of the analysis.

Lens: In the context of the present invention the term “lens” may beused as a component or a system that is enabled to spread or convergeacoustic energy. Any matter being able to influence the propagationcharacteristics of the generated acoustic energy shall be includedwithin the term “lens.”

Interface/Interface medium: In the context of the invention thepropagation path of the acoustic energy may comprise several componentslike the source, the full solid coupler and the cartridge. In order todescribe the transitions or areas where these different elements of thepropagation path get in physical contact to each other the termsinterface and interface medium are used. For example if a coupler isphysically contacted with the cartridge, the interface medium of thecoupler describes the material used in the coupler within this area ofthe coupler brought in contact with the cartridge.

Coupler: The term coupler will be used in the context of the inventionas an element that is part of the propagation path of the acousticenergy and transfers may be with other elements the acoustic energy fromthe source to the cartridge. Furthermore the term coupler will be usedsimilarly to the term full solid coupler.

Solid gel: In the context of the present invention “solid gel” comprisesa gel-forming material only. It is fully solid and it is at the sametime a gel. Liquid substances are fully avoided within a solid gel. Thuswater or hydrogel is avoided when using a solid gel. Thus the term “gel”is similarly used in the context of the invention to the term “solidgel.”

It should be noted that embodiments described in the following similarlypertain to the device for irradiating a sample with focused acousticenergy and the method for irradiating a sample with focused acousticenergy. Synergetic effects may arise from different combinations of theembodiments although they might not be described explicitly or indetail.

Further on, it shall be noted that all embodiments of the presentinvention concerning a method, may be carried out with the order of thesteps as described, nevertheless this has not to be the only andessential order of the steps of the method. All different orders andcombinations of the method steps are herewith disclosed.

According to a first aspect of the present invention, there is provideda full solid coupler for a complete dry coupling of acoustic energybetween a source and a cartridge. Accordingly, in a first exemplaryembodiment of the invention a device for irradiating a sample withfocused acoustic energy to treat the sample is presented, wherein thedevice comprises an instrument, a cartridge, a full solid coupler and asource for generating the acoustic energy. Furthermore the cartridge hasa chamber for receiving the sample and the full solid coupler provides acomplete dry coupling of the acoustic energy between the source and thecartridge. The instrument and the cartridge are adapted for insertingthe cartridge into the instrument wherein the cartridge and theinstrument are separable.

In the following possible further features and advantages of the deviceaccording to the first exemplary embodiment will be explained in detail.

In other words, by inserting the cartridge into the instrument acomplete dry propagation path for the focused acoustic energy from thesource to the sample is generated. AU different dry components of theinstrument, the cartridge, the full solid coupler and the source arethus connected in a complete dry manner after inserting the cartridgeinto the instrument. The coupler in general transmits the acousticenergy from one of its end to another. It shall explicitly be noted,that the full solid coupler is arranged at the device in such a way,that it complements or completes the propagation path of the acousticenergy between the source and the cartridge in a dry way. In other wordsthe propagation path comprises before insertion of the full solidcoupler a first dry partial propagation path and a third partialpropagation path. By inserting the coupler between these two parts, themissing second partial path is supplied. The complete propagation pathmay for example be formed firstly out of a material attached to afocusing transducer, secondly out of polymer based coupler and thirdlyout of a foil between the coupler and the cartridge. Thus a complete drycoupling between the source and the cartridge is achieved. Thus the fullsolid coupler does not have to form the whole propagation path byitself, but if it is desired, an exemplary embodiment of the inventionmay realize this.

Therefore, the use of water or hydrogel or any gel containing liquidsubstances is avoided. Thus after a completion of an irradiation of thesample with the acoustic energy a clear distinction between a possiblyleaking cartridge containing liquid matter and the coupling media can bemade. In other words, situations with a high contamination risk due tothe leakage of a cartridge may be recognized by a user of the devicemore clearly and even faster.

As the instrument and the cartridge are totally different componentsthat are physically separated or at least separable the volume of thesample to be treated can be chosen by selecting different cartridges.Furthermore the chamber of the cartridge may not be totally filled withthe sample and thus having an additional air layer within the chamberabove the sample. This may arise in several technical advantagescompared to so-called flow through systems. An exemplary advantage of anair layer above the sample is that with HiFu vigorous mixing could beintroduced, allowing treatment of sample volumes much larger than thefocal zone volume. For example by creating a fountain of the sampleliquid by means of HiFu irradiation a mixing mechanism via circulationof the sample liquid that is imminent in the fountain cycle can besupplied. Thereby the focal zone in which the HiFu energy creates thefountain may be quite small compared to the sample volume, butnevertheless a mixing process is initiated by HiFu via the fountain.Thus the need to irradiate the whole sample volume that is to be mixedmay be avoided by this exemplary embodiment of the invention. In otherwords a large sample may be treated by a relatively small device.

Additionally HiFu could create a fountain, which may be used to createcavitation at relative low (reduced) powers. The cavitation nuclei maybe introduced in the sample by the fountain droplets returning to theliquid which may reduce the power-threshold compared with homogenouscavity in water with an order of magnitude. In other words by creating afountain out of the sample (e.g. when the sample is a liquid) theminimum power for the transducer and thus the minimum acoustic energy tobe emitted from the source can be reduced. This may lead to advantagesdescribed in the context of the present invention.

In other words the fountain could in addition to cause a mixing in thesample and a reduction of cavitation power threshold be used for coolingthe sample, as the fountain creates much larger contact surface of thesample with the surrounding air within the cartridge.

The physical separation of the cartridge and the instrument may lead toa non-integrated system which means that the source, the coupler and thecartridge may be chosen and applied for a measurement independently fromeach other. In other words when the interface between the threeconstituting parts of the system (source, coupler and cartridge) isdefined an independent choice of those three constituting parts may bemade as long as the choice fits with the interface.

Because of the fact, that the size of the cartridge and the chamber areindependent from the size and shape of the source and of the coupler anenlargement of the cartridge volume is possible without having the needto change the acoustic characteristics of the device. A disadvantage offlow through systems compared to this embodiment of the invention may bethat an enlargement of the chamber is possible without having toincrease also the transducer.

Additionally it may be relied on focusing onto a focal zone and avoidinga dependency of an interaction of the acoustic energy with a wall of thechamber. In other words the walls of the chamber are not used as atransducer. In contrast to that known systems have to take into accountthat resonance frequencies of chamber walls are functions of geometryand material properties. These systems have to match that with thesource frequency. As it is not relied on interactions of the acousticfield and the walls in such a described way, an enlargement of thechamber may be done independently from the transducer choice.

As the cartridge is physically separable from the instrument thecartridge may be a disposable, consumable and removable cartridge whichmay lead to a cheap system for analyzing the sample with focusedacoustic energy. After a treatment of the sample the cartridge may bediscarded without having the need to discard the source or the coupler.Thus a plurality of measurements provided by one single instrument andone solid coupler and one source for a variety of different cartridgeswith different samples is a possible way using dry coupling.

The device may further comprise a lens for focusing the generatedacoustic energy onto the sample.

Furthermore the irradiation of the sample by focused acoustic energycauses a treatment of the sample.

The source or transducer could be a flat or curved piezo transduceroperating between kHz up to MHz frequencies. The diameter of thetransducer may be for example between 5 mm and 35 millimeters (mm) tofit with the volume range, for example between 0.2 milliliters (mL) and10 mL, one would like to process in the cartridge. The focal length ofthe transducers may vary from 5 mm to 80 mm. Transducer electric inputpower may vary from 2 watts (W) to 100 W. According to this exemplaryembodiment of the invention the treatment of samples is possible withlower powers compared to related known technology. Thus heating due toacoustic energy absorption of circumjacent matter, especially of thematter between the source and the sample is avoided, enabling theintroduction of dry coupling.

The transducer may operate in a continuous mode or in a burst mode.Applied signal to transducer could have different and varying forms:e.g. sinusoidal, block, triangular, or any combination thereof.Frequency may be additionally adjusted to compensate for frequency shiftto heating or to switch focal length.

The cartridge may have one of the following characteristics: disposable,consumable, removable, may contain one chamber or a lot of chambers, maycontain one sample or a lot of samples, industrial applicable. Thecartridge material is not limited to but may further for example bepolyethylene (PE), polypropylene (PP), polyethylene terephthalate (PET);polymethylpentene (PMP), Polymethylmethacrylate (PMMA), polycarbonate(PC) and polystyrene (PS).

In addition to that the cartridge is also a physically independentdevice from the coupler. Thus the cartridge is different and separablefrom the instrument and also from the coupler. This exemplary embodimentof the invention does not exclude, that the coupler is placed or fixedonto the cartridge or the instrument, but contains this possibility.

One main advantage is that all desired and needed processing of thesample may be done in one single chamber of the cartridge. Furthermorethe whole processing by the applied acoustic energy may be doneaccording to the sample-in result-out principle with all necessaryactuation coming from one single source of the device. By means of thefocused acoustic energy the sample may be treated with a lot ofdifferent functionalities like sample pretreatment and lysis in onesingle chamber being a process chamber. Especially HiFu may be used forthese processes.

To achieve a high intensity of the acoustic energy at the receivingposition (chamber in cartridge and thus at the sample) it is preferredthat the focus quality of the source or transducer and/or lens issufficient, that the acoustic attenuation of the materials in thepropagation path of the acoustic energy is sufficiently low which meansa low impedance and/or a low thickness, and that reflection at thematerial interfaces in the propagation path of the acoustic energy issufficiently low which means for the dry coupler that the thickness androughness of the two contacting layers should be sufficiently small.This exemplary embodiment of the invention meets these requirements.

Power may be supplied to the source from the instrument via e.g. leadsor brushes. The full solid coupler may comprise different pieces, partsor segments.

Furthermore the dry coupling may induce, that on the microscale thecontact between for example the source and the coupler (first layer)and/or the coupler and the cartridge (second layer) may approach directcontact condition, in other words as close as possible in order toachieve efficient dry coupling. Thus the surfaces of the two layers maybe on the microscale or nanoscale as conformal as possible to minimizeor eliminate air-pockets between the two layers in dry contact.

In other words to minimize or eliminate air pockets the followingrequirements may be met by the device: The surface roughness may besufficiently low of the source, the coupler, the cartridge, the fullsolid coupler and an interface medium. Also the used materials may besufficiently “flexible” to achieve conformality. Thereby a conformalityorder may be considered liquids>hydro gels>solid gels>rubbers>(elastic)foils>thermoplastic polymer>thermo harders, metals, ceramics and othersolid materials.

The acoustic energy or acoustic radiation may propagate through a firstpart of the path unfocused and may later be focused within a second partof the path to propagate focused through a third part of the path tillthe sample. Previous or subsequent focusing is also possible.

The required power for creating a cavitation process in the sample maybe reduced by this exemplary embodiment of the invention, as additionalnucleation sites may be introduced in the chamber (e.g. an element withan appropriate high surface roughness e.g. a rod) or a fountain may beinduced. Droplets falling back from the fountain into the sample mayreduce this power threshold. As the present construction enables bothpossibilities low power HiFu may be used for preparing and treating thesample.

As the required power may be decreased by the present invention,additional refraction, being generated at high intensities, may beavoided.

According to another exemplary embodiment of the invention the focusedacoustic energy is high intensity focused ultrasound (HiFu).

Thereby the source frequencies may be in the range of 0.2 MHz to 10 MHz,with amplitudes chosen to be sufficient efficient to create highpressure shock-waves and/or cavitation in the focal zone. Focal zonedimensions may be dependent on the source transducer type. Exemplarylength-scales for the indicated frequency range are (sub) millimeters.Furthermore flat or curved piezo transducer may be used operatingbetween 0.2 MHz and 10 MHz, or between 0.75 MHz and 3 MHz or between 1MHz and 2 MHz. The diameter of the transducer may be for example between5 mm and 35 mm to fit with the volume range (0.2 mL-10 mL) one wouldlike to process in the cartridge. The focal length of the transducersmay vary from 5 mm to 80 mm. Transducer electric input power may varyfrom 0.5 W to 100 W.

In other words, this exemplary embodiment of the invention may be usedas a HiFu molecular device for treating and/or analyzing molecularsamples. Thereby no liquid matter must be used for coupling the acousticenergy from the source to the sample. Thus liquid contamination risksmay be reduced and by using disposable or consumable cartridges anuncomplicated, cheap and fast way of measuring characteristics of thesample plus preparing the sample with the device and thus with HiFu maybe provided.

Due to the relatively short wavelength of HiFu compared to ultrasound,an enhanced focusing onto a smaller region is possible. This leads to aminiaturization advantage.

In addition to that various different focal region shapes may be usedfor treating the sample by the HiFu.

As HiFu enables a user to treat a sample e.g. with functionalities likemixing with a reagent, circulation, release of a cell, pathogen andmatrix from a swab, release of a cell, pathogen and matrix from a brush,liquefaction, incubation of the sample with a reagent at roomtemperature or elevated temperature, shaking, mixing; stirring,extraction, NA extraction, flow generation, sample homogenation,separating by centrifuging, and any combination thereof, lysis, lysis ofmicroorganisms, incubation of the sample with a reagent at room orelevated temperature, and any combination thereof a huge varietyapplications for the device is created.

Furthermore known systems may be limited dictated by physics becausereal miniaturization of the ultrasound transmitter may not be possible;known systems may thus be limited to about 100 mm. This embodiment ofthe invention may be miniaturized smaller than 100 mm.

Further on another disadvantage of known systems may be that theresonance frequency of the ultrasound chamber is design and materialdependent and should be matched with the chosen ultrasound transmitterfrequency. Manufacturing tolerances may have to include this dependency.In contrary to that, any resonance frequency of the device may not havebeen taken into account, as described above.

Furthermore other instruments using acoustic energy may be limited to asmall volume chamber as according to basic physical laws of mechanics anincreasing in dimensions means a reduction of the resonance frequency ofthe chamber or the system. Parallel existing requirements of theultrasound frequency originating in the specifications of the sample maythus make it useless to increase the size of the chamber. This may limitthe spectra of the applications of such a known instrument.

In contrary to that, a non integrated system is presented, in which thecartridge is physically independent i.e. separated from the source andthe coupler, as described above. It may be that no resonance frequencyof the chamber has to be taken into account, when selecting the desiredsize of the cartridge or the chamber. This is an important advantageabove the known technology.

Furthermore this exemplary embodiment is enabled to avoid, if necessary,a flow-through technique, which may complicate the combination withincubation at elevated temperature. In addition these flow-throughtechnologies may have the need to provide some kind of beads to thechamber. But in the case a flow through may be desired, the present ideais able to provide for that.

In other words this exemplary embodiment of the invention distinguishesfrom technologies using ultrasound impacting the wall of the chamber. Inthese known systems the resonance frequency is dependent from geometryand/or the material of the device.

Furthermore in contrast to flow-through systems which use homogenouscavitation, the power can be reduced in this exemplary embodiment, asthis exemplary embodiment of the invention may supply for an air layerin the chamber, which makes it possible to introduce nucleation sites orto produce a fountain as described above. By means of additionalnucleation sites like a rod that is introduced in the chamber or bymeans of the described fountain, the power threshold to initiatecavitation may be reduced. Furthermore it may be provided for anincubation possibility of the sample although not all of the samplefluid has to be in the focal zone.

This may enable the user to use smaller transducers and less power whichenables the introduction of the full solid coupler or dry coupling.Furthermore the combination of incubation may be facilitated.

In addition to that this exemplary embodiment of the invention is ableto use additional different functions, e.g. elution of swabs in aprocess chamber. As HiFu is used with dry coupling it allows to detectcartridge leakage and therefore contamination could be detected in anearly stage.

According to yet another embodiment of the present invention a source isone of part of the instrument or part of the cartridge.

In a first example of this embodiment the source may be implemented inthe instrument of the device. Thus a plurality of cartridges may beirradiated one after another by one and the same acoustic energy source.Thus measurement results of different cartridges may be more comparableand reliable as deviations originating from different sources can beexcluded.

In a second example of this embodiment of the invention the source ispart of the cartridge. For example, a cartridge may be provided with asource and a full solid coupler being situated between the source andthe cartridge. For example, they may be glued together to one unit. Alsoother fixation possibilities shall be comprised. By inserting this unitinto the instrument the electric connection between the power supply forthe source is plugged together. Thus a complete dry coupling isgenerated in this embodiment of the invention.

By means of the integration of the source into the cartridge apre-selection or pre-adaption of the specific source for certainmeasurement intentions is possible. Thus in combination with theinstrument different types of cartridges with specifically selectedsources for these cartridges and for specific measurements may be usedwith one single instrument. This means an increase operation field ofthe instrument. In addition to that the cartridges and the sources beingattached to the cartridges may be disposable and thus may provide acheap and uncomplicated solution for treating different samples indifferent cartridges with different sources attached by one singleinstrument.

According to another exemplary embodiment of the invention, theinstrument and the cartridge are arranged in combination in such a waythat by inserting the cartridge into the instrument the propagation pathfor transmitting the acoustic energy from the source to the sample isformed wherein the propagation path comprises only of non-fluidicmatter.

In other words, the interaction of the cartridge and the instrumentduring the insertion process yields to the complete dry couplingpropagation path. Therefore, the corresponding surfaces of theinstrument and the cartridge are brought together during an insertionprocess and they may be shaped in a for example form-closed way or in aforce-fit way. In addition to this shape fitting of the contour of theinstrument and the cartridge extra means for applying a pressure betweenthese elements and the additional coupler may be provided. In otherwords, only solid materials or gaseous materials, like air pockets, arepresent in the propagation path of the acoustic energy.

According to another exemplary embodiment of the invention, the fullsolid coupler is formed out of the material selected from the groupcomprising solid gel, rubber, elastic foil, polymer based material,thermoplastic polymers, polymer having a low acoustic attenuationcharacteristic, metal, semiconductor, ceramic, polypropylene, aluminum,and a stack of these materials.

It shall explicitly be noted, that the full solid coupler may be formedout of a polymer based material.

The used materials may obey elastic characteristics that allow aconformable adaption of the coupler to the shape of a component of thedevice for example of the cartridge or of the source. Thereby thematerial of the full solid coupler may be chosen in such a way, that airpockets at any interface within the propagation path are minimized oravoided to achieve efficient dry coupling. Furthermore the full solidcoupler may also contain the above mentioned materials as partialcomponents and other not mentioned materials may be contained in thefull solid coupler.

Calculations have shown that the stack could increase the amount ofenergy which could be transferred to the receiving position, but at theexpense of a more complicated coupler. In other words impedance matchingmay be used. Thus the full solid coupler may comprise severalcomponents, that together yield to a complete and efficient dry couplingof the acoustic energy from the source to the sample.

According to another exemplary embodiment of the invention, thecartridge comprises an acoustic window wherein the acoustic window ismade of a flexible foil and wherein the full solid coupler is physicallycontacted with the acoustic window by inserting the cartridge into theinstrument.

In order to achieve a high intensity of the focused acoustic energy atthe receiving position (chamber in the cartridge where the sample ispositioned) it may be essential that the attenuation of the materials inthe transport path of the HiFu is sufficiently low. Furthermore airpockets shall be minimized or eliminated by using surface roughness'sthat are sufficiently low. Also materials that are sufficiently flexibleto achieve conformity may be used. These requirements may be met by theacoustic window that is made out of a flexible material like a plasticfoil. Thereby the plastic foil may adapt its shape during an insertionof the cartridge into the instrument to the shape of the contact surfaceof the cartridge or the shape of the full solid coupler.

The acoustic window of the cartridge may be sufficiently large that thecross section of the HiFu cone at the chosen acoustic window distancefits completely in the window. The acoustic window could be flat orcurved. The acoustic window is of made of a thin layer of a lowattenuation polymer, e.g. PP, PMP. It is also important that theremaining part of walls of the lysis chamber below the fluid level maybe sufficiently thin to reduce acoustic losses and to limit heating ofthe chamber housing

According to another exemplary embodiment a contact pressure between thefull solid coupler and the cartridge is applied, wherein the contactpressure is generated by at least one method from the group comprisingapplying over pressure in the chamber of the cartridge, applying localunder pressure outside of the cartridge, and pressing the cartridge andthe full solid coupler against each other by means of a force.

The contact pressure between the solid coupler and the cartridge surfaceis applied in a sufficient way to get rid of air or air pockets at theinterface or at any intermediate layer in the propagation path of theacoustic energy. Pressing e.g. a convex shaped solid coupler against aflat cartridge, with the cartridge material being sufficiently flexibleto become conformal to the shape of the solid coupler may be possiblesolution. In addition to that the coupler may also have such aflexibility.

Another exemplary embodiment may be a dry interface solution comprisinga smooth spherical or conical shaped HiFu transducer and a flexiblecartridge foil.

Thereby the contact pressure yields to a force that presses a least thethree components source, coupler and cartridge together in such a way,that air pockets may be minimized between some or all intermediatecontacting surfaces. Therefore especially smooth and flexible materialsmay be used for these surfaces.

According to another exemplary embodiment of the invention, the fullsolid coupler has a first contact surface for contacting the acousticwindow and the cartridge has a second contact surface for contacting theacoustic window. Furthermore at least one of the first contact surface,the second contact surface and the acoustic window has a surfaceroughness value selected from the group comprising smaller than 0.5micrometers (nm), smaller than 1 μm, and smaller than 2 μm.

Due to this embodiment of the invention air pockets and thustransmission losses in the propagating acoustic energy may be minimizedor eliminated.

An interface medium between the instrument and cartridge to enableacoustic energy transport across dry interface may be made of a lowattenuation material like rubber (e.g. RT 615), (elastic) foil (e.g. PP,PP based Thermoplastic Elastomer, PMP), or thermoplastic polymer (e.g.PP). The interface layer could be part of the instrument or of thecartridge. For example the cartridge bottom layer contacting the couplercould also be at the same time the interface medium.

According to another exemplary embodiment of the invention, thepropagation path has a gradient of acoustic impedance, wherein thegradient is monotonously decreasing in a direction from the source tothe sample.

This embodiment may lead to further reduction of acoustic energy losses,as a coupling from one component to another component in the propagationpath may be improved due to the gradient of the acoustic impedance. Byapplying such an acoustic impedance profile within the propagation pathreflection and absorption of the focused acoustic energy may be reduced.This may lead to a better yield or spoil of a given power.

The acoustic impedance of the materials used within the propagation pathof the acoustic energy is going from relative high on the side of thesource to relative low at the sample/cartridge site. In addition to thatprinciple laws of acoustics may be used to optimize the choices ofdimensions and material of the device and its components.

According to another exemplary embodiment of the invention, the fullsolid coupler is selected from the group comprising a coupler being aphysically separate component placed between the source and thecartridge, a coupler being part of the source, a coupler being part ofthe cartridge and any combination thereof.

For example, a configuration with the source being a piezo transducercombined with a metal lens on top that has a polymer coupler on top ofthe metal lens is possible. Also a curved source working simultaneouslyas a lens may be provided with a polymer coupler on top of that curvedsource. The coupler may physically be bonded to the source or thecartridge but it can also be hold on top of one of these components byexternal pressure applied to these components. Referring now to thefollowing FIGS. 10 to 14 a large variety of combinations of arrangingand fixing the coupler between the source and the cartridge arepossible. Placing the coupler on the source, on the cartridge, on alens, on a second additional coupler and on an acoustic window withdifferent fixation possibilities like pressing together, gluingtogether, depositing a coupler on a component, and any combinationthereof are comprised within this embodiment of the invention.

According to another exemplary embodiment of the invention, a lens forfocusing the generated acoustic energy onto the sample is furthercomprised. Thereby the lens is selected from the group comprising a lensbeing a physically separate component placed between the source and thecartridge, a lens being part of the source, a source with a focusingshape being the lens, an array of sources that yield to a focus acousticenergy, a lens being part of the cartridge, a lens made out of apolymer, having a low acoustic attenuation characteristic, a metal lens,a ceramic lens, a polypropylene lens, an aluminum lens, a hybrid lensand any combination thereof.

Lens may be made of a low attenuation polymer, metal or ceramic. Forenvironmental reasons the lens may be integrated in the consumable andmade of a polymer, e.g. PP.

As a first characteristic of the lens the lens is able to focus thegenerated acoustic energy onto the sample. In order to reduce thetransmission losses the lens may be attached to the source. For examplea metal lens may be fixed onto a piezo transducer yielding to theemission of a focused acoustic field. Furthermore an array of aplurality of sources may be spatially placed in such a way andelectronically driven in such a way that the superposition of all thesingular acoustic fields yields a focused acoustic field. Furthermore itis possible that the lens is part of the cartridge for example beingfixed to the bottom of the cartridge. In addition to that this examplemay further comprise a source being part of the cartridge.

In order to create multi-focality also a hybrid lens may be used in thisexemplary embodiment of the invention. Thereby the lens has at least twodifferent emitting zones which means, that the different emitting zonesof the lens deviate from each other by at least one of the followingcomponents shape, surface roughness, material, and any combinationthereof. To shortly summarize the function of a hybrid lens it has to besaid that an incoming homogeneous acoustic field will be transferred bythe hybrid lens into a non-homogenous acoustic field having for exampletwo different focal regions.

According to another exemplary embodiment of the invention, in the onesingle chamber of the cartridge pretreatment and lysis are applied tothe sample by means of the focused acoustic energy. Thereby pretreatmentis a method selected from the group comprising mixing with a reagent,circulation, release of a cell, pathogen and matrix from a swap, releaseof a cell, pathogen and matrix from a brush, liquefaction, incubation ofthe sample with a reagent and/or enzyme at room temperature or elevatedtemperature, shaking, mixing, stirring, extraction, NA extraction, flowgeneration, sample homogenation, separating by centrifuging and anycombination thereof. Furthermore lysis is a method selected from thegroup comprising mixing with a reagent, mixing with a reagent differentto the reagent applied during pretreatment, circulation, lysis ofmicroorganisms, incubation of the sample with a reagent at room orelevated temperature or a temperature different from the temperatureapplied during pretreatment and any combination thereof.

It shall explicitly be noted that this combination of pretreatment andlysis in one single chamber by means of the focused acoustic energyoriginating from only one single source may be applied without providinga dry coupling. No full solid coupler or a propagation path out ofcompletely dry media is necessary.

Accordingly, a second aspect of the present invention is directed to theapplication of pretreatment and lysis to the sample by means of focusedacoustic energy in the one single chamber of the cartridge, i.e. inparticular in the same chamber. An exemplary embodiment of this aspectof the present invention provides a device for irradiating a sample withfocused acoustic energy to treat the sample, the device comprising aninstrument, a cartridge, and a source for generating the acousticenergy. The cartridge has a chamber for receiving the sample. Theinstrument and the cartridge are adapted for inserting the cartridgeinto the instrument. The cartridge and the instrument are separable. Thedevice is designed such that pretreatment and lysis are applicable tothe sample in the chamber of the cartridge by means of the focusedacoustic energy.

In addition to that an exemplary embodiment of this second aspect of thepresent invention further relates to a corresponding instrument forirradiating a sample with focused acoustic energy to treat the sample,the instrument comprising a source for generating the acoustic energy.The instrument is adapted to receive a cartridge being separable fromthe instrument, the cartridge providing a chamber for receiving thesample. The instrument is designed such that, when the cartridge isbeing inserted in the instrument, pretreatment and lysis are applicableto the sample in the chamber of the cartridge by means of the focusedacoustic energy.

Correspondingly, a cartridge is provided in another exemplary embodimentwhich cartridge for an instrument for irradiating a sample with focusedacoustic energy generated by a source to treat the sample comprises achamber for receiving the sample. The cartridge is adapted for beinginserted into an instrument and being separable from the instrument. Thecartridge is designed such that when being inserted into the instrumentpretreatment and lysis are applicable to the sample in the chamber bymeans of the focused acoustic energy.

In addition to that exemplary embodiments of this aspect of theinvention further relate to a corresponding method for pre-treating andlysing a sample in one single chamber by means of focused acousticenergy like for example HiFu originating from one single source,preferably by such device, and a computer program element characterizedby being adapted when being used to control a device for pre-treatingand lysing a sample to cause the device for performing the steps of thiscorresponding method.

These exemplary embodiments may for example combine pretreatment andlysis in a single chamber by using single focus HiFu, but also the useof multi focus HiFu is possible. But also any combination withincubation is possible.

In other words, manual steps for doing sample pretreatment can beavoided by an exemplary embodiment of the device, the correspondinginstrument and cartridge, the corresponding method and the computerprogram element. Pretreatment is integrated in the cartridge to increaseease-of-use and to decrease fluidic interfacing with the external worldand contamination risk. Furthermore pretreatment and lysis functions areintegrated in one single chamber that could be exposed to HiFu and/orheated and/or cooled to reduce complexity, costs and size of the deviceand the procedure for doing treatment and lysis together. Pretreatmentand lysis functions advantageously are processed without the sampleleaving the chamber in between, and/or advantageously are processed in afully automated manner, and/or advantageously are processed sequentiallyor simultaneously.

This second aspect of the invention may be used for any applicationrequiring pretreatment and/or lysis. Applications may not be limited tohealthcare, life science, food industries and veterinary practice. Thisrelates to any embodiment of the invention.

Especially for lysing difficult micro-organisms the state of the arttechnique of applying thermal lysis has several insufficiencies. Incontrary to that this aspect of the present invention uses focusedacoustic energy, especially HiFu for solving these problems. By means ofsuch a fully integrated in vitro preparation and detection instrument asystem for sample-in result-out tests is provided, especially fornucleic acid (NA), protein or cell detection. Furthermore nucleic acidanalysis, protein analysis and cell analysis may be possible by a socalled micro total analysis system.

Additionally, existing lysing methods comprise grinding or bead beatingwhich may be avoided here.

In general nucleic acid sample preparation protocols are morecomplicated than cell or protein preparation protocols. Although thisaspects of the invention may be for the major part on nucleic acidsample preparation it is not limited to this.

For this reason a single solution with a high flexibility is needed toaccommodate these deviations in needed pretreatment. This aspect of theinvention meets these requirements with high degree of flexibility onpretreatment and lysis protocols.

It shall be noted that preferred embodiments of other aspects of thepresent invention shall be considered as preferred and disclosedembodiments with respect to the present aspect, too, and vice versa.

According to another exemplary embodiment of the invention, the deviceis adapted in such a way that it generates at least two different focalregions at the sample.

It shall explicitly be noted that this exemplary embodiment of theinvention may be applied or implemented without having the need toprovide for the complete dry coupling features. In other words, thecreation of a multi-focality by the device may also be used incombination with non-solid coupling matter.

Accordingly, a third aspect of the present invention is directed to thegeneration of two different focal regions at the sample. In an exemplaryembodiment of this third aspect of the present invention a device ispresented for irradiating a sample with focused acoustic energy to treatthe sample comprising an instrument, a cartridge, and a source forgenerating the acoustic energy. The cartridge has a chamber forreceiving the sample. The instrument and the cartridge are adapted forinserting the cartridge into the instrument. The cartridge and theinstrument are separable. The device is designed for generating at leasttwo different focal regions of acoustic energy at the sample.

In addition to that an exemplary embodiment of the third aspect of thepresent invention further relates to a corresponding instrument forirradiating a sample with focused acoustic energy to treat the sample,the instrument comprising a source for generating the acoustic energy.The instrument is adapted to receive a cartridge being separable fromthe instrument and providing a chamber for receiving the sample. Theinstrument is designed for generating at least two different focalregions of acoustic energy at the sample when the cartridge is insertedin the instrument.

Correspondingly, a cartridge is provided in another exemplary embodimentwhich cartridge for an instrument for irradiating a sample with focusedacoustic energy generated by a source to treat the sample comprises achamber for receiving the sample. The cartridge is adapted for beinginserted into an instrument and being separable from the instrument. Thecartridge is designed for allowing generating at least two differentfocal regions of acoustic energy at the sample when being inserted inthe instrument.

Furthermore it shall explicitly be noted that a corresponding method forgenerating at least two different focal regions at the sample by thedevice and a corresponding computer program element for controlling adevice generating a multi-focality to the sample is comprised withinthis embodiment. Thereby the computer program element may becharacterized by being adapted when in use on a device for creating amulti-focality to the sample to cause the device for performing thesteps of the corresponding method.

In other words, a treatment protocol using two different focal zones forproviding different focus conditions is provided. For example, focusconditions for doing mixing a liquid circulation by means of focusedacoustic energy may be different from the requirements for doing lysiswith for example microorganisms. This embodiment of the invention meetsthese requirements.

By providing at least two different focal regions at the sample thedevice may provide for attractive simple and cheap molecular diagnostictests. Furthermore complex arrangements of piezo arrays, complicatedsystems and/or drivers can be avoided by this exemplary embodiment ofthe invention. Furthermore the integration of several differentfunctionalities (like e.g. mixing, circulating, and lysing) into onechamber that are processed by the two different focal regions aminiaturization of the molecular diagnostic device is possible.

In other words the molecular diagnostic device is a multi-focality HiFumolecular diagnostic device for applying different focal regions to thesample. This can be used for generating and combining differenttreatment functionalities. For example point-like focused HiFu may beoptimal for doing lysis and zone-like focused HiFu may be optimal formixing and/or circulating. Thereby point like means a comparativelysmall focal region, and a zone-like focus means a comparatively largefocal region. Different focal regions may also differ in shape and insize. These different focal requirements are met by this exemplaryembodiment of the invention.

Furthermore lysis by means of HiFu requires high acoustic pressures.High pressures are achieved by good quality focusing which is achievedby this exemplary embodiment of the invention by means of a first highlyfocused part of the generated acoustic energy. In contrary to that torelease particles or cells from swabs, to release and homogenize fecesfrom a carrier, e.g. swabs, brush, to homogenize liquid present in thechamber with reagents added to the chamber mixing and circulation may berequired in one single chamber of the cartridge. Thus a second part ofthe generated acoustic energy is focused to a comparatively large,zone-like second focal region at the sample. Thus two differenttreatment functionalities may be applied to the sample during the sametime, in one single chamber and without any manual intervention of auser.

According to another exemplary embodiment of the invention, the at leasttwo different focal regions are generated by means of an elementselected from the group comprising a plurality of sources, a singlesource and a hybrid lens, one single source with different roughnesszones and one single source being excited differently at differentpositions of the source, and any combination thereof. Such element maybe embodied as an element external to the cartridge, or an elementbelonging to the cartridge, or as an element integrated into thecartridge.

A plurality of sources comprises at least two single sources, as well asan array of sources being electronically controlled in such a way thatthe superposition field of all the sources yields to a total fieldhaving at least two focal regions. Furthermore the hybrid lens mayconsist of a moderately focusing material and a highly focusingmaterial. These materials may be positioned at different parts of thelens yielding to multi-focality. For example a concave shaped hybridlens may be attached to a flat source like a transducer. But also acurved transducer with a curved hybrid lens made out of a moderatelyfocusing material and a highly focusing material is possible. In orderto find an optimized distribution of these two different materialsacoustic modeling may be performed on different configurations. Forexample, the lens may be formed out of polypropylene. Furthermore lensradius may vary due to the application of the device. In order to createa multi-focality at the receiving position where the sample is located,the source may also be provided with different roughness zones, whichmeans that the surface of the source obeys different surface roughnessvalues.

The different emitting zones, more detailed the respective surfaces ofthese zones, may have different roughness properties. These differentroughness properties yield to different acoustic irradiatingcharacteristics of the zones, which leads to at least two differentfocal zones. Thereby the source or transducer itself may have thesezones. But also an additional component may be added on top of thetransducer, wherein the component obeys these different surfaceroughness characteristics. In other words the gist of this possibilityis that the surface of the transducer is segmented in a smooth and rougharea delivering respectively highly and moderately focused acousticenergy, especially HiFu, to the sample.

It shall be noted that preferred embodiments of other aspects of thepresent invention shall be considered as preferred and disclosedembodiments with respect to the present aspect, too, and vice versa.

According to another exemplary embodiment of the invention, the focusedacoustic energy is used for reducing the viscosity of the sample.

It shall explicitly be noted that this embodiment of the invention doesnot necessarily need to contain all the dry coupling features. Inparticular no full solid coupler or a completely dry propagation path isnecessary.

Accordingly, a fourth aspect of the present invention is directed tousing the focused energy for reducing the viscosity of the sample. In anexemplary embodiment of this fourth aspect of the present invention adevice is provided for irradiating a sample with focused acoustic energyto treat the sample comprising an instrument, a cartridge, and a sourcefor generating the acoustic energy. The cartridge has a chamber forreceiving the sample. The instrument and the cartridge are adapted forinserting the cartridge into the instrument. The cartridge and theinstrument are separable. The device is designed for using the focusedacoustic energy for reducing the viscosity of the sample.

In addition to that an exemplary embodiment of the fourth aspect of thepresent invention further relates to a corresponding instrument forirradiating a sample with focused acoustic energy to treat the sample,the instrument comprising a source for generating the acoustic energy.The instrument is adapted to receive a cartridge being separable fromthe instrument and providing a chamber for receiving the sample. Theinstrument is designed for using the focused acoustic energy forreducing the viscosity of the sample when the cartridge is inserted intothe instrument.

Correspondingly, a cartridge is provided in another exemplary embodimentwhich cartridge for an instrument for irradiating a sample with focusedacoustic energy generated by a source to treat the sample comprises achamber for receiving the sample. The cartridge is adapted for beinginserted into an instrument and being separable from the instrument. Thecartridge is designed for allowing reducing the viscosity of the sampleby means of focused acoustic energy applied to the sample when beinginserted into the instrument.

In addition to that an exemplary embodiment comprises a correspondingmethod for reducing the viscosity of the sample, preferably by suchdevice, and a corresponding computer program element. Thereby thecomputer program element is characterized by being adapted when in useon a device for reducing the viscosity of the sample by means ofirradiating the sample with focusing acoustic energy to cause the devicefor performing the steps of the corresponding method.

In order to reduce the viscosity of a sample like for example BAL,sputum, blood, feces, or any other sample present on a swab thisembodiment of the invention suggests to use focused acoustic energy forexample HiFu to cause this reduction. This method may be implemented ina complete sample-in result-out solution in which a subsequentpretreatment and lysis of the sample may be possible in the one chamberof the cartridge. Thus by means of only one single source a completeprocess of viscosity reduction, further pretreatment and lysis ispossible.

For example, a source having the following characteristics may be usedfor the reduction of the sample viscosity. 3.0 MHz transducer with adiameter of 25 mm a focal length of 22 mm. Furthermore the bottom of thecartridge may be set at 15 mm distance of the transducer. Exemplarypower of 5 W may be applied to the sample for approximately 300 s. Bymeans of such a HiFu application the sample may be more homogeneousafter such a HiFu exposure and the viscosity may drop from the originalviscosity to for example a water-like viscosity. Thus it can beconcluded that HiFu forces combine the ability to reduce the molecularweight of the macromolecules and as a result the viscosity and theability to circulate and mix a sample in a process chamber.

It shall explicitly be noted that this exemplary embodiment of theinvention may be used for any application requiring circulation and/ormixing in the sub-millimeter volume range in a device. The applicationsmay be also in the life sciences, lab-on-the-chip, and mTASapplications.

It shall be noted that preferred embodiments of other aspects of thepresent invention shall be considered as preferred and disclosedembodiments with respect to the present aspect, too, and vice versa.

According to another exemplary embodiment of the invention, a detectionunit for applying measurements on the sample is further comprised.Thereby the irradiation of the sample with the focused acoustic energyleads to a treatment of the sample.

In other words, this exemplary embodiment of the invention provides fora complete sample-in result-out system, where no manual step has to bedone by the user. A sample may be inserted into the device and by meansof the focused acoustic energy the sample is treated in a desired way.Subsequently or also previously measurements may be applied to thesample by means of the detection unit. Thereby the device is enabled todeliver the measurement results to for example a user interface to theuser. For example, functionalities like liquefaction, stirring, mixing,circulation, pretreatment, incubation and lysis may be done before orafter any measurement of the detection unit by means of the focusedacoustic energy. A fully automated system is thus provided to the user.

It shall further be noted that this exemplary embodiment of theinvention may not necessarily contain all dry coupling features. Inparticular, no full solid and dry coupler or a completely drypropagation path is necessary.

Accordingly, a fifth aspect of the present invention is directed to adetection unit for applying measurements on the sample. In an exemplaryembodiment of this fifth aspect of the present invention a device isprovided for irradiating a sample with focused acoustic energy to treatthe sample comprising an instrument, a cartridge, and a source forgenerating the acoustic energy. The cartridge has a chamber forreceiving the sample. The instrument and the cartridge are adapted forinserting the cartridge into the instrument. The cartridge and theinstrument are separable. The device comprises a detection unit forapplying measurements on the sample.

In addition to that this exemplary embodiment of the invention furtherrelates to a corresponding instrument for irradiating a sample withfocused acoustic energy to treat the sample, the instrument comprising asource for generating the acoustic energy. The instrument is adapted toreceive a cartridge being separable from the instrument and providing achamber for receiving the sample. The instrument comprises a detectionunit for applying measurements on the sample when the cartridge isinserted into the instrument.

Correspondingly, a cartridge is claimed in another exemplary embodimentwhich cartridge for an instrument for irradiating a sample with focusedacoustic energy generated by a source to treat the sample comprises achamber for receiving the sample. The cartridge is adapted for beinginserted into an instrument and being separable from the instrument. Thecartridge is designed such that when being inserted into the instrumenta detection unit may apply measurements on the sample.

In addition to that it shall be noted that this exemplary embodimentcomprises a corresponding method for applying measurements on the sampleby such device and a corresponding computer program element. Thereby thecomputer program element is characterized by being adapted when in useon such a sample-in result-out system to cause the device for performingthe steps of the corresponding method.

This allows in vitro treatment of the sample by means of e.g. HiFu andat the same time in vitro detection which leads to a complete sample-inresult-out system.

Especially for a molecular device being a device according to anembodiment of the invention and which device is enabled to extract,purify, amplify and detect nucleic acids it shall be stated thefollowing: Extraction and/or purification of nucleic acids is based onadsorption and/or desorption on a solid surface. Any surface offeringsufficient capture area should be regarded as a part of an embodiment ofthe invention. Common surface capture embodiments are (for examplemagnetic) particles and membranes. Any capturing material capable ofdelivering nucleic acids of sufficient quality for multiplicationpurposes should be regarded as part of an embodiment of the invention.Widely used materials are e.g. silica, magnetized silica, iron-oxide,aminogroup functionalized polystyrene. Also other materials arepossible.

The detection unit and thus the detection method of choice may bedependent on the application area like e.g. nucleic acids, protein orcell detection.

For nucleic acid amplification and detection e.g. a large number ofisothermal and thermal cycling amplification methods are described.Polymerase chain reaction (PCR) is one of the most used methods. Thesample-in result-out system according to this exemplary embodiment ofthe invention implements such a PCR functionality into the chamber wherethe sample is also treated by means of HiFu.

PCR is further subdivided in two subcategories namely end-point andreal-time PCR (rtPCR). Of these two rtPCR is most widely used (rtPCRamplification is running in parallel with detection). For detection ofnucleic acids one may for example use detectable markers such asfluorescent markers which may be incorporated in the amplified nucleicacids during PCR. Other detectable labels or even label-free methods mayalso be used.

For protein detection, common approaches such as a combination ofantibody capture and optical readout, e.g. fluorescence, of magneticreadout may be used.

For cell detection, optical methods as they are widely used to count,analyze cell shape, etc, but (di) electrophoretic and electricalproperties could also used to detect/characterize cells.

All the before mentioned detection possibilities of this embodiment ofthe invention correspond to the detection unit that is used in thisembodiment of the invention. Thus the realized sample-in result-outsystem may incorporate any of these detection or measurement features.

According to another exemplary embodiment of the invention, thedetection unit is for applying at least one measurement to the sampleselected from the group comprising optical measurements, magneticmeasurements, thermal measurements, electrical measurements, chemicalmeasurements, sonic measurements, and any combination thereof.

The device may further comprise at least one of: an extraction unit; anucleic acid amplification unit; a reagent storage unit; a detectionunit a detection unit for applying measurements on the sample whereinthe detection unit is for applying at least one measurement to thesample selected from the group comprising optical measurements, magneticmeasurements, thermal measurements, electrical measurements, chemicalmeasurements, sonic measurements, and any combination thereof. Accordingto this embodiment the apparatus may comprise, for instance: anextraction unit; an extraction unit and a nucleic acid amplificationunit; an extraction unit, a nucleic acid amplification unit, and adetection unit. In each of these options a reagent storage unit may bepresent in addition to the elements of each option listed in theprevious sentence. The extraction unit allows a nucleic acid to beobtained from a sample processed by the apparatus. The nucleic acidamplification unit allows a nucleic acid obtained from the sample to beamplified (using, for instance, PCR). The reagent storage unit comprisesa reagent needed for, for instance, extraction and/or amplification.

In order to have a wide spectrum of measurement possibilities differenttypes of sensors and detectors may be installed within the device.Additionally it may be advantageous to combine the already existingultrasonic means for actuating or treating the sample with thepossibility to do sonic measurements. The detection unit may be alsopart of the cartridge. In other words optical readout, but also otherdetection labels e.g. magnetic, electrical, electro-magnetic especiallyradio-frequency applied techniques but also labelless methods arepossible.

According to another exemplary embodiment of the invention, the devicefurther comprises a processor for coordinating a treatment protocol, adata processor, a display and a user interface.

It shall be noted that preferred embodiments of other aspects of thepresent invention shall be considered as preferred and disclosedembodiments with respect to the present aspect, too, and vice versa.

According to another exemplary embodiment of the invention the fullsolid coupler is made out of a polymer based material; and wherein thepolymer based material has a glass transition temperature T_(g) selectedfrom the group comprising: T_(g)≧−30° C.; T_(g)≧−10° C.; T_(g)≧−5° C.;T_(g)≧20° C.; T_(g)≧40° C.; T_(g)≧60° C.; T_(g)≧80° C.; T_(g)≧100° C.;T_(g)≧120° C.; T_(g)≧130° C.; T_(g)≧140° C.; T_(g)≧150° C.; andT_(g)≧160° C.

It shall be noted, that the relevance of the glass transitiontemperature of the material of the full solid coupler gets moreimportant the higher the intensity of the HiFu is. For low intensity,for example when the input power P of the transducer is smaller than 3Watt the value of T_(g) may not be that relevant. This may be seen inFIG. 22. Medium intensity, P being e.g. between 3 and 6 Watt, selfenforced attenuation as described above and hereinafter may play a moreserious role which may require a polymer with a sufficiently high T_(g).At high intensities above e.g. 6 Watt the relevance of the choice of thepolymer based on his T_(g) value even gets more important.

It may be necessary that at high intensity HiFu applications at roomtemperature the T_(g) may have to be above room temperature(approximately 50° C.).

It has been found that materials with relative high glass transitiontemperature T_(g) keep contrary to lower T_(g) materials duringoperation of the device and thus during the acoustic energy transmissiontheir low attenuation characteristics. Thus an application of theselow-attenuation high-T_(g) materials as full solid couplers enables veryefficient transmission of ultrasound intensities relevant for e.g.treatment of samples, e.g. lysis of cells. Especially for HiFuapplications as defined above this is an advantageous effect realized bythe invention. In other words by using these materials a reduced powerprovided to the source may be necessary to realize a certain HiFu powerin the focal region. Thus treatment and/or pretreatment functionalitiesmay be realized with a reduced power value. This may save energy andcosts. In other words the effect of self-enforced attenuation of thecoupling material may be avoided by the invention. The attenuation permeter of the propagation path may thus be reduced.

In order to provide for a better understanding of this exemplaryembodiment of the invention the following description of the physicalprocesses shall be noted:

Intrinsic to attenuation is that the coupler material temperature maystart increasing. Further the attenuation of acoustic energy may alsoincrease in parallel. This exemplary embodiment of the invention nowprovides for materials that have the advantage to keep a relatively lowattenuation even when their temperature starts increasing during e.g.HiFu operation in the MHz range.

Examples for such materials may be polypropylene with T_(g)approximately −18° C., epoxy with T_(g) approximately 60° C. andsilicons with T_(g) approximately 60° C., approximately 100° C. andapproximately 125° C.

It has to be noted that a sufficiently high glass temperature is relatedto the attenuation at the start of the test, the ultrasound intensity,the thermal conductivity of the setup (transport of heat generated) andthe exposure time.

In other words the choice of the polymer with a certain T_(g) valuedepends on several parameters like the attenuation value of the polymerat the beginning of the HiFu transmission through the polymer as fullsolid coupler and thus before any absorption or heat generation hasstarted. Furthermore the applied intensity or the power of the sourcedetermines that choice of the polymer. Additionally the thermalconductivity of the surrounding of the coupler is a parameter whichinfluences the choice of a polymer with a sufficiently high T_(g) value.A high thermal conductivity of the system around the coupler results inslower temperature rise and lower maximum temperature, if HiFu issufficiently long exposed to reach equilibrium.

Especially for relatively high-intensity ultrasound applications thismay be particularly relevant. For example when doing lysis with the HiFuenergy within the sample the necessary power may be relatively high.Thus for lysing methods using HiFu this exemplary embodiment may bequite advantageous.

In other words the full solid coupler present in the propagation betweenthe source and the destination being the cartridge has a reducedattenuation of the acoustic energy. In addition to that tuned or matchedimpedances of the materials transmitting the acoustic energy may beused, to minimize reflection losses when passing material interfaces.

Thus the device provides for a complete dry coupling with the possiblefollowing advantages: ease-of-use for the operator and reduction of testturn-around-time, as a consequence other applications run byless-skilled personal could be envisioned.

Polymers are a particular advantage class of materials to be used ascouplers, because of the rich variety of materials available, the shapeand dimensional design freedom, easy replication and associated relativelow costs. This may further be described in detail in FIGS. 21 to 23.

According to another exemplary embodiment of the invention wherein thepolymer based material has been cured at a curing temperature T_(c)selected from the group comprising: T_(c)≧20° C.; T_(c)≧40° C.;T_(c)≧60° C.; T_(c)≧70° C.; T_(c)≧80° C.; T_(c)≧90° C.; T_(c)≧100° C.;T_(c)≧110° C.; T_(c)≧120° C.; T_(c)≧130° C.; ≧140° C.; T_(c)≧150° C.;T_(c)≧160° C.; T_(c)≧170° C.; and T_(c)≧180° C.

The attenuation of the full solid coupler may further be reduced ceterisparibus when the curing temperature of the polymer based material duringpolymer fabrication is increased. This may further be described indetail in FIGS. 21 to 23.

It has been found, that during the polymer fabrication, which includes acuring process step, the curing temperature during that curing processstep at least partially determines the transition glass temperature ofthe built polymer material. As described above a sufficiently high T_(g)value has certain advantages for applications in a HiFu moleculardevice. Thus by defining the curing temperature to a certain value adesired T_(g) value may be realized in the polymer. Such a process stepmay be part of a method according to another exemplary embodiment of theinvention.

According to another exemplary embodiment of the invention, a method forirradiating a sample with focused acoustic energy to treat the sample isprovided. Thereby the method comprises the following steps: providingfor an instrument, providing for a cartridge, providing for a full solidcoupler, providing for a source for generating the acoustic energy, andinserting the cartridge into the instrument. Furthermore the cartridgehas a chamber for receiving the samples and due to the inserting of thecartridge into the instrument a complete dry coupling of the acousticenergy between the source and the cartridge is provided. The cartridgeand the instrument are separable.

According to another embodiment of the invention an instrument forirradiating a sample with focused acoustic energy to treat the sample ispresented. The instrument comprises a source for generating the acousticenergy, a full solid coupler, wherein the instrument is adapted toreceive a cartridge containing the sample, wherein the full solidcoupler provides a complete dry coupling of the acoustic energy betweenthe source and the cartridge, when the cartridge is inserted in theinstrument; wherein the cartridge and the instrument are separable andwherein the instrument and the cartridge form a device according to oneof the above described embodiments.

This embodiment of the invention may be used with HiFu acoustic energyin order to treat the sample with methods or functionalities like mixingand/or lysing in e.g. one single chamber. Furthermore the instrument maycomprise a detector and an excitation source that may both be for doingoptical, electrical, magnetic and/or mechanical measurements.Additionally a lens may be comprised in the instrument.

In other words the dry coupling may be realized by the full solidcoupler that is part of the instrument. Before the presence of thecartridge a complete dry propagation path from the source through thefull soil coupler is realized. By inserting the cartridge into theinstrument the whole dry propagation path between the source and thesample is completed and the acoustic energy may be transferred to thesample in order to treat the sample.

According to another embodiment of the invention cartridge for aninstrument for irradiating a sample with focused acoustic energy totreat the sample is presented, the cartridge comprising a chamber forreceiving the sample, a full solid coupler, wherein the cartridge isadapted for being inserted in the instrument. Furthermore the full solidcoupler provides a complete dry coupling of the acoustic energy betweenthe source and the cartridge when the cartridge is inserted in theinstrument, wherein the cartridge and the instrument are separable andwherein the instrument and the cartridge form a device according to oneof the above described embodiments.

The full solid coupler may be permanently fixed to the cartridge. Butother solutions are possible. The source may for example be part of theinstrument. By inserting the cartridge into the instrument the fullsolid propagation path between the source and the sample is established.

Furthermore the source may be comprised by the cartridge. Thus byinserting the cartridge into the instrument electrical leads from theinstrument are contacted with the source, in order provide the sourcewith electrical energy.

For the two before mentioned embodiments it shall explicitly be noted,that the full solid coupler is arranged at the instrument or at thecartridge in such a way, that the full solid coupler does not have toform the whole propagation path by itself and other additional drycoupling elements may be present. Nevertheless if it is desired anexemplary embodiment of the invention may realize this.

It shall further be noted that a computing unit may be part of theinstrument. It may be a separate unit in communication with theinstrument, or computing tasks may be distributed over computer unit andinstrument.

It shall further be noted that all computer program elements mentionedabove as exemplary embodiments of the invention might be stored on acomputing unit, which might also be part of an embodiment of the presentinvention. This computing unit may be adapted to perform or induce theperforming of the steps of the method described above. Moreover, it maybe adapted to operate the components of the above-described device. Thecomputing unit can be adapted to operate automatically and/or to executethe orders of a user. Furthermore the computing unit can request theselection from a user to process the input from the user.

The embodiments concerning computer program elements cover both acomputer program, that right from the beginning uses the computerprogram element and a computer program that by an update turns anexisting program into a program that uses the invention.

According to a further embodiment of the present invention, acomputer-readable medium is presented wherein the computer-readablemedium has a computer program element stored on it which computerprogram element is described by the preceding or following sections.

It may be seen as a gist of the invention that a consumable cartridgebeing separable from the instrument generates a complete dry couplingpropagation path for focused acoustic energy when the cartridge isinserted into the instrument. Thereby the dry coupling reaches from thesource generating the acoustic energy to the sample.

It has to be noted that some of the embodiments of the invention aredescribed with reference to different subject-matters. In particular,some embodiments are described with reference to method type claimswhereas other embodiments are described with reference to apparatus typeclaims. However, a person skilled in the art will gather from the aboveand the following description that unless other notified in addition toany combination or features belonging to one type of subject-matter alsoany combination between features relating to different subject-mattersis considered to be disclosed within this application.

The aspects defined above and further aspects, features and advantagesof the present invention can also be derived from the examples ofembodiments to be described hereinafter and are explained with referenceto examples of embodiments. The invention will be described in moredetail hereinafter with reference to examples of embodiments but towhich the invention is not limited.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a schematic image of a device for irradiating a sample withfocused acoustic energy to treat the sample according to an exemplaryembodiment of the present invention.

FIG. 2 shows a schematic image of a cartridge having an acoustic windowaccording to an exemplary embodiment of the present invention.

FIG. 3 shows a schematic image of a cartridge according to an exemplaryembodiment of the present invention.

FIGS. 4 to 8 show schematic images of sources of a device according toan exemplary embodiment of the invention.

FIG. 9 shows a schematic image of several components of an instrumentaccording to an exemplary embodiment of the present invention.

FIGS. 10 to 14 show exemplary overviews of possible configurations of adevice according to exemplary embodiments of the present invention.

FIG. 15 shows a schematic image of electronic components being used fora device according to an exemplary embodiment of the present invention.

FIG. 16 shows a treatment protocol that is processed by a deviceaccording to an exemplary embodiment of the present invention.

FIGS. 17 to 19 show schematic images of devices generatingmulti-focality to the sample according to an exemplary embodiment of thepresent invention.

FIG. 20 shows a flow diagram representing a method according to anexemplary embodiment of the present invention.

FIG. 21 shows a schematic image of a device for irradiating a samplewith focused acoustic energy to treat the sample according to anexemplary embodiment of the present invention.

FIGS. 22 and 23 show a diagrams of results obtained with a device forirradiating a sample with focused acoustic energy to treat the sampleaccording to an exemplary embodiment of the present invention.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Similar or relating components in the several figures are provided withthe same reference numerals. The view in the figure is schematic and notfully scaled.

FIG. 1 shows a device 100 for irradiating a sample 101 with focusedacoustic energy to treat the sample according to an exemplary embodimentof the present invention. It can clearly be seen that the device hasseveral components being an instrument 102, a cartridge 103, and asource 105 (shown only with dashed lines) for generating the acousticenergy. Furthermore, a schematic drawing of the propagation path 106(dashed dotted lines) of the acoustic energy starting at the source 105and ending at the sample 101. Thereby the cartridge has a chamber 110for receiving the sample 101. Inside of the shown instrument 102 a fullsolid coupler (not shown) 104 is provided in order to generate apropagation path without non-fluidic matter. Thereby the source 105 andthe full solid coupler 104 are located inside of the instrument 102 andthus cannot directly be seen. Furthermore the instrument 102 and thecartridge 103 are adapted for inserting the cartridge into theinstrument wherein the cartridge and the instrument are separable. Itshall be noted that the hidden components like the source, the lens, thefull solid coupler and the acoustic window can be seen on the followingFIG. 9 showing an exploded view and FIG. 2 respectively.

Additionally a detection unit 111, e.g. a sensor, is shown inside of thecartridge in order to do measurements on the sample after or before apossible treatment by the focused acoustic energy. Furthermore aprocessor for coordinating a treatment protocol 112 is shown which islinked with the detection unit 111 and which is also connected to adisplay 114 and a data processor 113. The processor 112 for coordinatinga treatment protocol is connected to the device 100 and is furtherconnected to the detection unit 111. Thus the processor 112 is enabledto control this complete-in result-out system in which in a fullyautomated way a treatment of a sample by means of focused acousticenergy especially by HiFu can be combined with analysis and measurementsas for example optical measurements, magnetic measurements, thermalmeasurements, electrical measurements, chemical measurements, sonicmeasurements and any combinations thereof.

Due to the use of HiFu and the corresponding short wavelength (comparedfor example to known ultrasound applications operating in the 20 kHz-100kHz range) the size of focal region can be decreased and thus aminiaturization of the whole molecular device is possible. This is ahighly important advantage of the shown embodiment of the presentinvention with for example hospital or lab requirements to have realsmall size systems because of the very limited space available in thesesurroundings. Furthermore, the combination of the functionalitiestreatment, pretreatment, lysis and previously or subsequently donemeasurements may reduce the costs and time of such a sample treating ormolecular diagnosis.

Additionally it may be possible to provide as such a device 100 with amulti-focality setup. Thereby the device generates at least twodifferent focal regions at the sample 101. This may be done by at leasttwo different sources, a single source and a hybrid lens, or a singlesource with different roughness stones. Furthermore a combination ofthese possibilities is also possible.

Furthermore this device 101 may be used to reduce the viscosity of asample by means of the focused acoustic energy especially by using HiFu.

In addition to that the device makes it possible to combine in onesingle chamber 110 pretreatment and/or incubation and/or lysis by meansof focused acoustic energy originating from only one single source 105.Especially a HiFu application is possible. Thereby pretreatment andlysis' may comprise different functionalities that have been describedin previous sections. This may reduce costs and time of such a sampletreating or molecular diagnosis and also the space claimed of the devicemay be reduced due to the integration of both functionalities into onechamber. Furthermore the technical complexity of the device may bereduced.

A pretreatment method or a lysis method may be processed or carried outby means of the focused acoustic energy, especially by HiFu and thus bythe acoustic source or transducer generating the HiFu spot at theposition of the sample yielding to a pretreatment and/or lysis of thesample. But also other devices that may be integrated into the moleculardiagnostic device and that are necessary to carry out the method maygenerate the desired method. For example an additional heating device,cooling devices, or reagent applicator (dispenser) with supply lines maybe integrated in the molecular diagnostic device to cause incubationwith an additional reagent at elevated temperature.

A reagent may for example be lysozyme enzyme which may first be mixedand subsequently incubated at 37° C. Especially mixing, circulation,liquefaction and homogenation may be done by means of the irradiation ofthe sample with HiFu.

Furthermore also lysis of micro-organisms like e.g. gram-negative andgram-positive bacteria, fungi and yeast may be done by means of HiFuwith the device 100 shown in FIG. 1. Lysing may further compriseincubation of the sample with a reagent at room temperature or elevatedtemperature. Reagents may for example be GuHCl/prot K which is firstmixed and subsequently incubated at approximately 56° C. and optionallycooled down to environmental temperature or GuSCN which is first mixedand subsequently incubated at approximately 70° C. and optionally cooleddown to approximately 25° C.

Optionally the chamber has at its outlet a filter or in its outletchannel a filter to assure that debris is not transported to theextraction functionality of the cartridge.

FIG. 2 shows an acoustic window 107 of the cartridge 103 wherein theacoustic window is made of a flexible material which is shown as aplastic foil 108. It can be seen that the circular-shaped acousticwindow 107 that is shown in a bottom view is covered by the plastic foil108 being the interface medium that may adapt itself to the shape offirstly the cartridge 103 and secondly to a full solid coupler or sourcemay be brought in contact with the plastic foil directly on the shownsurface 108. 115 shows the bottom part of the cartridge on which aflexible foil is e.g. laser welded.

FIG. 3 shows the cartridge 103 with the chamber 110 in its normal orworking orientation which is a 180° rotation compared to FIG. 2. Inother words, FIG. 2 shows the bottom part 115 of the cartridge with itsbottom side and FIG. 3 shows the cartridge with the bottom part 115 fromthe upper side. The shown cartridge and foil clamp can then together asone unit be inserted into the device 100 of FIG. 1 and can be pushed ontop of the instrument 102. This inserting process will form apropagation path for transmitting the acoustic energy from the source105 (shown with dotted lines) in FIG. 1 to the sample 101 in FIG. 1.

FIG. 4 shows an example of a possible source used in a device accordingto an exemplary embodiment wherein a source 105 and a coupler 104 isshown wherein the here shown example is a polymer coupler.

FIG. 5 shows another example of a source creating the focused acousticenergy especially HiFu wherein the source 105 may be a piezo transducerand a metal lens 109 is fixed on top of that for example flattransducer. Additionally a coupler 104 is provided for example a polymercoupler.

In contrary to that FIG. 6 shows a polymer coupler configuration inwhich a curved source 105 is combined with a polymer coupler 104. Inaddition to that for example a lens may be located on top of the polymercoupler being provided with another for example polymer coupler on topof the lens to provide for an efficient dry coupling towards thecartridge.

FIG. 7 shows a piezo configuration in which a flat piezo transducerworking as a natural focusing source 105 can be seen. Additionally avery thin polymer layer is applied to modify the roughness of thesurface to promote efficient dry coupling. In addition to that theelectric leads are also shown.

FIG. 8 shows another possible configuration of the source components inwhich a metal lens 109 is directly contacted to the flat transducerworking as a source 105. As will be later on seen in FIGS. 10 to 14 anycombination of these configurations is possible which leads to a widespectrum of applications.

FIG. 9 shows an exploded view of an instrument 102 comprising a heatsink 900, different housing rings 901 partially building up the housingfor the full solid coupler 104 that might e.g. be a polymer basedmaterial or a solid gel, an additional ring 902. Furthermore the source105 is shown as a piezo transducer. Additionally the full solid coupler104 is denoted with dotted lines. These elements may be part of theinstrument 102 and they may build a receiving component that byinserting a cartridge on top of the foil clamp 903 creates a propagationpath that only consists of non-fluidic matter. The elements 901, 902 and903 are part of the housing of the coupler, too. The housing is madesuch that the height of the coupler could be modified by choosing numberof housing rings 901. The foil clamp 903 is clamped to the foil (notdepicted) which is used to cover the coupler.

FIG. 10 shows an overview of combinations of possibilities to create drycoupling. Thereby the first row gives information about the setup of thecartridge 103, the second row gives information about the setup of thecoupler 104, the third row gives information about the setup of the lens109 and the fourth row gives information about the setup of the sourceor transducer 105. It can be seen that five different configurations areshown as examples. 1001 shows a solid gel coupler configuration wherein1002 shows a metal lens polymer coupler configuration and 1003 describesa polymer coupler configuration. 1004 describes a solution for the drycoupling where a piezo only configuration (wherein the piezo has a thinpolymer layer to modify the roughness surface of the transducer) is usedand 1005 describes how a metal lens configuration may be set up in orderto reach dry coupling. 1001 shows that a source may be shaped like alens and thus defines the generation and the focusing of the acousticenergy. Furthermore shown in column 1002 the lens may be physicallycombined for example may be glued together with the solid coupler 104.Furthermore the full solid coupler 104 may be directly attached to thesource 105 as shown in column 1003. But also a direct contact betweenthe cartridge and the piezo source is possible as shown in 1004.Additionally the metal lens configuration describes that at a curvedshaped source 105 can be attached a biconcave shaped lens e.g. a metallens.

Other setup possibilities may be shown in the detailed overviews 1100within the FIGS. 11, 12, 13 and 14. These overviews are more detailedthan FIG. 10 because two additional rows are inserted in order todistinguish between the fact whether a component is part of thecartridge, is part of the source (which means is part of the instrument)or is a physically separated component.

It shall explicitly be noted that any shown and described component maybe part of the transducer, of the cartridge or may be a physicallyseparated component. In addition to that any combination of componentsmay be used in order to separate different functionalities. For examplea thin foil, having a high flexibility may be used to adapt the shape ofa transducer. In combination with a full solid coupler having lessflexibility but lower attenuation than the foil, this corresponds to theseparation of the functionalities attenuation and flexibility. This maylead to an advantageous combination of different components to achieveefficient dry coupling.

Row 1101 describes, if there is an entry, that the full solid coupler ispart of the cartridge. In contrary to that 1102 describes the fact thatthe full solid coupler is part of the source and thus part of theinstrument. Also both possibilities may be arranged at an devicesimultaneously. As a third possibility 1104 describes that the fullsolid coupler is a physically separated component being inserted intothe propagation path. Again it can be seen that a combination of lensand source 1103 may be provided. As can be seen from FIGS. 11 to 14 ahuge variety of setup possibilities for the dry coupling of the deviceusing for example HiFu is possible.

FIG. 15 shows exemplary electronic components 1500 being used togenerate the focused acoustic energy. Thereby a possible functiongenerator, a power amplifier, a scope and an ultrasonic transducer areconnected together in order to create the acoustic field. After havingfocused the emitted acoustic energy it impinges the sample and causesdifferent sono-chemical or sono-physical reactions. This is thetreatment of the sample caused by device. In other words FIG. 15 shows aconfiguration of a lab setup to generate and investigate the setupperformance. An industrial device may not include a scope and thefunction generator and the amplifier may be embodied in specific andcustom made electronics.

FIG. 16 shows a possible treatment protocol for applying pretreatmentand lysis in one single chamber by only one single source. Treatmentprotocol 1600 has several steps for example the protocol starts with aHiFu pretreatment of the sample 1603, subsequently a mixing 1604 isapplied to the sample wherein afterwards an incubation with differentmatter 1605 is possible. Subsequent additional mixing and incubationsteps are possible. These different functionalities created or caused bythe acoustic energy due to sono-chemical or sono-physical interactionsare all part of the pretreatment 1601. Subsequently a lysis 1602 ispossible within the same single chamber and can be caused by the samesingle source that has been processed the pretreatment. As possiblesteps mixing and incubations may be mentioned. But also special HiFulysis 1606 and additional filter steps 1607 are possible. Therebyreference sign 1608 describes any sample with a target material to bedetected, e.g. feces, blood, urine, sputum, BAL, CSF, tissue, swab orbrush. Furthermore a first pretreatment reagent (e.g. chemicalcompound(s) and/or enzyme(s)) is shown with 1609. A second pretreatmentreagent (chemical compound(s) and/or enzyme(s) is shown with referencesign 1610 and 1611 depicts a third pretreatment reagent (chemicalcompound(s) and/or enzyme(s)). A first lysis reagent (chemicalcompound(s) and/or enzyme(s)) is shown by 1612. 1613 shows an extractionreagent, e.g. to prepare for DNA binding on silica. The shown figure isonly an exemplary embodiment and a filter does not have to be inside thelysis chamber.

FIG. 17 shows a multi-focality setup 1700 of the device according toanother exemplary embodiment of the invention. It can be seen that thecartridge 103 having a chamber 110 with a sample 101 also features thepossibility to have an air volume 1701 above the sample. Furthermore twodifferent sources 105 are applied in the setup in order to generate afirst focal region 1702 and 1703 showing a second focal region.Furthermore the acoustic window of the cartridge should have a lowattenuation and minimal thickness to avoid heating of the material andto realize a high intensity in the focal regions. For mass production aninjection moldable polymer is preferred. It may be preferred that nocontact is made of the focal regions with the walls of the chamber. Athigh intensities this may result in melting of the wall. It may furtherbe desired that the transducer with the large focal zone 1702 is placedopposite to the air volume 1701. This results in optimal mixing andcirculation and may have lower risk on melting the chamber wall.

FIGS. 18 a and 18 b show multi-focality of the device working forexample in the HiFu range, may be generated by only one single source.Thereby FIG. 18 a shows a multi-focality setup 1700 with a hybrid lens1800 having a first emitting zone 1801 and a second emitting zone 1802and a third emitting zone 1803. It is also possible that in a concentricsetup the first and the third emitting zones are equal. It can furtherbe seen, that in the sample 101 three different focal regions 1804 to1806 are generated. In a concentric setup it is thus the case that 1804and 1806 describe the same focal region having a ring-like shape aroundthe second focal region 1805.

It can be seen that the source 105 may be of a flat shape and the hybridlens 1800 is attached to the source.

FIG. 18 b shows a multi-focality setup 1700 wherein the hybrid lens 1800has got a shape that is adapted to the shape of the curved source 1500.In FIG. 18 b the hybrid lens has three emitting zones and three focalregions originating from the three emitting zones. The differentemitting zones may consist of different focusing material. For example,the outer material forming zone 1801 and 1803 may be of moderatelyfocusing outer material wherein the inner material forming the zone 1802may be a highly focusing material. The segmented lens 1800 thuscomprises highly focusing material and moderately focusing material.This may be the case for FIG. 18 b. These different focal regions mayenable a user to process different functionalities like mixing andlysing simultaneously by only using one single source. This may reducethe times of for example a molecular test and furthermore costs andspace requirements may be reduced as only one single source is needed.Additionally technical problems and maintenance costs are reducible.

Thereby the distribution of the differently focusing materials can beadapted the desired treatment, lysis or analyzing application. Thus nospecific material distribution within the hybrid lens or segmented lensis excluded by this exemplary embodiment of the invention.

The following paragraph relates to modeling of a combination comprisinga flat transducer and a curved lens to verify the hybrid lens concept. Apossible setup may be for example a high impedance material like forexample aluminum, a low impedance material like polypropylene taken as alens material, a lens radius and internal diameter of the chamber likefor example 8 mm and polypropylene is taken as chamber wall materialwith a thickness of 0.5 mm, a fluid height is 35 mm and the frequencyfor the modeling is 1 MHz and prescribed pressure piezo is 1.000 Pa. Theresults of the modeling disclosed that the maximum pressure along thecentral axis of symmetry remains at a very constant high level whengoing from a complete high impedance material (aluminum) to increasingsegment sizes of low impedance material like polypropylene. In otherwords, the pressure remains at the level sufficient high to obtainlysis. Secondly the results revealed that a minimum and maximum pressureconditions are created outside the central axis of the chamber when thepolypropylene segment size is sufficiently large to create mixing.Effective working of the hybrid construction is achieved when the highindex material (aluminum for instance) is typically between ⅕ and ½ ofthe total lens when a low index material is a low dissipation plastic.Thus a hybrid lens is an option to generate multifocal acoustic energyespecially multifocal HiFu from a single piezo element. This solutioncould be used for HiFu across dry interfaces as well as for liquid orhydrogel coupling and direct contact with the fluid.

FIG. 19 shows a multifocal setup 1700 wherein a source 105 has differentsurface roughness zones. 1903 shows a top view of the circular source105 having a first surface roughness zone 1904 and a second surfaceroughness zone 1905 yielding to multi-focality. It can be seen that thefirst focal region 1900 and the second focal region 1901 are differentfrom each other. Here the third focal region 1902 is the same as thefirst focal region 1900 because the second roughness zone 1905 is aring-shaped surface that yields to a ring-shaped focal region 1900 and1902 around the second focal region 1901. Due to different roughnessesof the surfaces a different coupling to material transmitting theacoustic energy is given. Therefore, different roughnesses yield indifferent focal regions.

It shall explicitly be noted that the multi-focality due to differentsurface roughnesses may not be used with the dry coupling features ofthe present invention and may be applied independently on a device forirradiating a sample with multi focused acoustic energy to treat thesample.

For example, in the range of 1 to 2 MHz the effect may be moderate for aroughness of 10 μm and may be significantly higher for a roughness of50-80 μm. Thus, a curved transducer with rough and smooth segments is anoption to generate multifocal HiFu from a single piezo element. Comparedto different solutions with lenses or a plurality of sources thisembodiment may be simpler.

FIG. 20 shows a flow diagram describing a method for irradiating asample with focused acoustic energy to treat the sample wherein thefollowing steps are comprised and for an instrument S1, providing for acartridge S2, providing for a full solid coupler S3, providing for asource for generating the acoustic energy S4. Furthermore inserting thecartridge into the instrument S5 wherein the cartridge has a chamber forreceiving the sample and wherein due to the inserting of the cartridgeinto the instrument a complete dry coupling of the acoustic energybetween the source and the cartridge is provided. Furthermore thecartridge and the instrument are separable.

FIG. 21 shows a schematic drawing of an instrument device comprising atransducer 105, a full solid coupler 104, a cartridge 103 having achamber 110 for a sample to be treated with e.g. HiFu by the instrument102. The bottom 2100 of the cartridge has an acoustic window made out ofa foil.

FIG. 22 shows a diagram 2200 in which the advantages of a full solidcoupler with a sufficiently high glass transition temperature T_(g) areillustrated. It can be seen from the graphs 2203-2207, that a full solidcoupler with higher glass transition temperature T_(g) provides for lessattenuation of the ultra sound energy within the full solid coupler.These results shall be describe in detail hereinafter.

The x-coordinate 2201 depicts the input power that is provided to thesource 105 (not shown) which generates the acoustic energy e.g. theHiFu. The y coordinate depicts the so called clipping time. This is thetime between the source generating e.g. the HiFu is switched on and thecomplete disappearance of the fountain (clipping). This fountaingeneration has been described above. It is created by the HiFu waves andis used to reduce the power threshold at which cavitation in the samplesets in. The fountain is consisting of the sample material (e.g. aliquid). As the generation of such a fountain depends on the acousticenergy that is transmitted through the full solid coupler to the samplethe disappearance of the sample means a reduction of transmittedacoustic energy. Different materials with different glass transitiontemperatures are observed within the test of the results shown in FIG.22.

In other words clipping is taken as a measure for the development of theattenuation or absorption with time of the observed full solid couplermaterial. Results for a variety of materials and thicknesses arepresented in FIGS. 22 and 23.

Thereby FIG. 22 shows results from a 3 mm thick silicon 601 coupler 2203having a glass transition temperature of 60° C. 2204 depicts the resultsfrom a 3 mm thick full solid coupler made out of epotek 301 having aglass transition temperature T_(g) of approximately 60° C. 2205 depictsthe results of a 6 mm thick silicon 601 coupler having a glasstransition temperature of 60° C. 2206 depicts the results of a fullsolid coupler that is 1 mm thick and made out of polypropylene (PP)having a glass transition temperature T_(g) of approximately −18° C.,2207 depicts the results of a full solid coupler made out of 5 mm thickepotek 301 having a glass transition temperature of approximately 60° C.All examples have cure temperatures of 60 C except PP.

FIG. 22 shows that PP is even at moderate intensity a rather poorperformer. Attenuation of both epoxy and silicone increases as expectedwith thickness of the full solid coupler. Attenuation of epoxy is forsilicon lower than for epoxy. For all of these high T_(g) materialsclipping is observed for continuous input power<6 Watt. This power maybe insufficient for sample treatment. Thus for broad treatmentpossibilities of a molecular diagnostic device the invention providesfor sufficiently high T_(g) polymers.

Additional experiments have disclosed that firstly the observedphenomena is not due to a change over time of the transducer. Secondlythe effect may be reversible (if the material is not exposed toburn-through intensities). After about 1 min the material is returned toits original state and the experiment could be repeated. Thisobservation suggests a temperature-material property relationship.

FIG. 23 shows a diagram 2300 in which the effect of the curingtemperature of the polymer based material used as a full solid coupleris shown. X-coordinate 2301 depicts the input power and y-coordinate2302 depicts the time to failure i.e. the clipping time. 2303 to 2306depict the graphs of the different full solid coupler. 2303 depicts theresult of a full solid coupler with the curing temperature T_(c) is 100°C., 2304 depicts T_(c) is 125° C., 2305 depicts T_(c) is 60° C. and 2306also depicts the results of a coupler with T_(c) is 60° C. In otherwords FIG. 23 shows that the effect attenuation is also dependent on thecuring temperature. With increasing curing temperature the clipping timeincreases significantly. An exemplary embodiment of the invention usesthis advantage. In other words in general a higher curing temperaturesT_(c) directly translates into a higher glass transition temperatureT_(g).

Additional experiments with the material cured at 60° C. have shownthat:

Firstly fountain disappeared if water of 80° C. or more is used.Secondly with a duty cycle of 20% the clipping time shift to >120seconds for peak power between 0 W and 65 W (average power 13 W). For apeak power of 90 W (average power 16 W) the clipping time has reduced to10 seconds.

Possible exemplary equipment devices for these test may be thefollowing: PM5193 Programmable Synthesizer/function generator 0.1 mHz-50MHz, Amplifier: ENI 240L Power Amplifier 50 dB 20 kHz-10 MHz or ARworldwide KAA204 RF Power Amplifier 50 dB 0.5-100 MHz 200 W, TektronixTDS3014: Four Channel Color Digital Phosphor Oscilloscope; Agilent4395A: 10 Hz-500 MHz/10 Hz-500 MHz/10 kHz-500 MHzNetwork/Spectrum/Impedance Analyzer and HiFu piezo transducer: JR20/60supplied by Dongfang Jinrong.

In the claims the word “comprising” does not exclude other elements orsteps, and the indefinite article “a” or “an” does not exclude aplurality. Reference signs shall not limit the scope of the claims.

LIST OF REFERENCE NUMERALS

-   100 Device-   101 Sample-   102 Instrument-   103 Cartridge-   104 Full solid coupler-   105 Source-   106 Propagation path-   107 Acoustic window-   108 Flexible material-   109 Lens-   110 Chamber-   111 Detection unit-   112 Processor for coordinating a treatment protocol-   113 Data processor-   114 Display-   115 Bottom part-   900 Heat sink-   901 Housing rings-   902 Additional ring-   903 Foil clamp-   1000 Overview of combination possibilities to create dry coupling-   1001 Solid gel coupler configuration-   1002 Metal lens polymer coupler configuration-   1003 Polymer coupler configuration-   1004 Piezo only configuration (wherein the piezo has a thin polymer    layer to modify roughness surface)-   1005 Metal lens configuration-   1100 Detailed overviews of combination possibilities to create dry    coupling-   1101 Row describing that the full solid coupler is part of the    cartridge-   1102 Row describing that the full solid coupler is part of the    instrument-   1103 Component combining the functionality of a lens and a source    (curved source)-   1104 Row describing that the full solid coupler is a physically    separated component-   1500 Electronics being used to generate the focused acoustic energy-   1600 Possible treatment protocol for applying pretreatment,    incubation and lysis in one single chamber by one single source-   1601 Pretreatment part of the protocol-   1602 Lysis part of the protocol-   1603 HiFu pretreatment-   1604 Mixing-   1605 Incubation-   1606 HiFu lysis-   1607 Filtering-   1608 Sample with a target material to be detected-   1609 First pretreatment reagent-   1610 Second pretreatment reagent-   1611 Third pretreatment reagent-   1612 First lysis reagent-   1613 Extraction reagent-   1700 Multi-focality setup-   1701 Air volume above the sample-   1702 First focal region-   1703 Second focal region-   1800 Hybrid lens-   1801 First emitting zone of the hybrid lens-   1802 Second emitting zone of the hybrid lens-   1803 Third emitting zone of the hybrid lens-   1900 First focal region-   1901 Second focal region-   1902 Third focal region-   1903 Top view of source 105 with different roughnesses zones-   1904 First roughness zone of the source-   1905 Second roughness zone of the source-   S1 Providing for an instrument-   S2 Providing for a cartridge-   S3 Providing for a full solid coupler-   S4 Providing for a source for generating the acoustic energy-   S5 Inserting the cartridge into instrument

Having described the invention, the following is claimed:
 1. A devicefor irradiating a sample with focused acoustic energy to treat thesample, the device comprising: an instrument; a cartridge having achamber for receiving the sample; a full solid coupler; and a source forgenerating acoustic energy, wherein the full solid coupler is configuredto provide a complete dry coupling of the acoustic energy between thesource and the cartridge, wherein the instrument and the cartridge areadapted for inserting the cartridge into the instrument, and wherein thecartridge and the instrument are separable.
 2. The device according toclaim 1, wherein the source is configured so that the acoustic energy ishigh intensity focused ultra sound (HiFu).
 3. The device according toclaim 1, wherein the instrument and the cartridge are arranged incombination in such a way, that by inserting the cartridge into theinstrument a propagation path for transmitting the acoustic energy fromthe source to the sample is formed, and wherein the propagation pathconsists only of non-fluidic matter.
 4. The device according to claim 1,wherein the full solid coupler comprises a material selected from thegroup consisting of solid gel, rubber, elastic foil, polymer basedmaterial, thermoplastic polymers, polymer having a low acousticattenuation characteristic, metal, semiconductor, ceramic,polypropylene, aluminum, and a stack of these materials.
 5. The deviceaccording to claim 1, wherein the cartridge comprises an acoustic windowmade of a flexible material, and wherein the full solid coupler isphysically contacted with the acoustic window by inserting the cartridgeinto the instrument.
 6. The device according to claim 5, wherein thefull solid coupler has a first contact surface for contacting theacoustic window, the cartridge has a second contact surface forcontacting the acoustic window, and at least one of the first contactsurface, the second contact surface and the acoustic window has asurface roughness value selected from the group consisting of smallerthan 0.5 micrometers, smaller than 1 micrometers, and smaller than 2micrometers.
 7. The device according to claim 3, wherein the propagationpath has a gradient of an acoustic impedance that is monotonouslydecreasing in a direction from the source to the sample.
 8. The deviceaccording to claim 1, further comprising a lens for focusing theacoustic energy onto the sample, wherein the lens is selected from thegroup consisting of a lens being a physically separate component placedbetween the source and the cartridge, a lens being part of the source, asource with a focusing shape being the lens, an array of sources thatyield to focused acoustic energy, a lens being part of the cartridge, alens made out of a polymer having a low acoustic attenuationcharacteristic, a metal lens, a ceramic lens, a polypropylene lens, analuminum lens, a hybrid lens, and any combination thereof.
 9. The deviceaccording to claim 1, wherein in one single chamber of the cartridgepretreatment and lysis are applied to the sample by the acoustic energy;wherein the pretreatment is a method selected from the group consistingof mixing with a reagent, circulation, release of a cell, pathogen andmatrix from a swab, release of a cell, pathogen and matrix from a brush,liquefaction, incubation of the sample with a reagent at roomtemperature or elevated temperature, shaking, mixing; stirring,extraction, NA extraction, flow generation, sample homogenation,separating by centrifuging, and any combination thereof, and whereinlysis is a method selected from the group consisting of mixing with areagent, circulation, lysis of microorganisms, incubation of the samplewith a reagent at room or elevated temperature, and any combinationthereof.
 10. The device according to claim 1, wherein the device isadapted in such a way, that it generates at least two different focalregions at the sample.
 11. The device according to claim 10, wherein theat least two different focal regions are generated by an elementselected from the group consisting of a plurality of sources, one singlesource and a hybrid lens, one single source with different roughnesszones, and one single source being excited differently at differentpositions of the source, and any combination thereof.
 12. The deviceaccording to claim 1, further comprising at least one of: an extractionunit; a nucleic acid amplification unit; a reagent storage unit; adetection unit a detection unit for applying measurements on the samplewherein the detection unit is for applying at least one measurement tothe sample selected from the group consisting of optical measurements,magnetic measurements, thermal measurements, electrical measurements,chemical measurements, sonic measurements, and any combination thereof,wherein the irradiation of the sample with the acoustic energy leads toa treatment of the sample.
 13. The device according to claim 1, whereinthe full solid coupler is made out of a polymer based material having aglass transition temperature T_(g) selected from the group consistingof: T_(g)≧−30° C.; T_(g)≧−10° C.; T_(g)≧−5° C.; T_(g)≧20° C.; T_(g)≧40°C.; T_(g)≧60° C.; T_(g)≧80° C.; T_(g)≧100° C.; T_(g)≧120° C.; T_(g)≧130°C.; T_(g)≧140° C.; T_(g)≧150° C.; and T_(g)≧160° C.
 14. The deviceaccording to claim 13, wherein the polymer based material has been curedat a curing temperature T_(c) selected from the group consisting of:T_(c)≧20° C.; T_(c)≧40° C.; T_(c)≧60° C.; T_(c)≧70° C.; T_(c)≧80° C.;T_(c)≧90° C.; T_(c)≧100° C.; T_(c)≧110° C.; T_(c)≧120° C.; T_(c)≧130°C.; T_(c)≧140° C.; T_(c)≧150° C.; T_(c)≧160° C.; T_(c)≧170° C.; andT_(c)≧180° C.
 15. A cartridge for an instrument for irradiating a samplewith focused acoustic energy generated by a source to treat the sample,the cartridge comprising: a chamber for receiving the sample; and a fullsolid coupler, wherein the cartridge is adapted for being inserted inthe instrument and for being separable from the instrument, and whereinthe full solid coupler provides a complete dry coupling of the acousticenergy between the source and the cartridge when the cartridge isinserted in the instrument.
 16. A method of irradiating a sample withfocused acoustic energy to treat the sample, the method comprising thefollowing steps: providing a cartridge with a full solid coupler;placing the sample in a chamber of the cartridge; inserting thecartridge into an instrument having a source for generating acousticenergy so that a complete dry coupling of the acoustic energy betweenthe source and the cartridge is provided, wherein the cartridge and theinstrument are separable.